The present invention relates to methods and compositions for inactivating viruses present in samples/process streams of biological origin.
It is desirable to use biological materials as sources for medicinal and industrial intermediates and products. Due to the very nature of the biological materials or their methods of production, biological materials may contain unwanted agents of viral origin that may be pathological or otherwise undesirable. The intended end-use of materials derived from biological sources may require a reduction in the biological activity of viral agents known to be present, or that are potentially present, in the source material or process additives.
Reduction of viral activity in materials is commonly accomplished by a number of techniques including the use of heat, steam, pressure, chemical treatments and other methods. However, these techniques may irreversibly alter the properties of the biological source material or the desired substances to be obtained from same. In such cases, gentle, non-denaturative and specific methods are required to reduce the biological activity of viruses without damaging the desired molecules or substances of interest.
Prior methods known in the art for inactivation of viruses in labile process streams include photochemical treatments in the presence of Psoralens, solvent-detergent treatments (U.S. Pat. No. 4,481,189), caprylic acid treatments (U.S. Pat. No. 4,939,176), the use of UVC radiation (Vitex Technologies, formerly NY Blood Center), ultra short time heating (Charm Technologies, charmbio.com), photodynamic inactivation in presence of phenothiazine dyes (U.S. Pat. No. 4,534,972), and the use of low-molecular-weight electrophilic agents that bind to nucleic acids (Vitex, Inactine product 4 patents).
The present invention relates to methods and processes of inactivating a viral contaminants in a biological source material or process intermediate. The process of the present invention involves contacting the biological source material (e.g., a host cell, cell supernatant, cell lysate, blood plasma, tissue homogenate, or other biological materials) containing a biomolecule (e.g., a recombinant or native protein, lipid, nucleic acid, or carbohydrate) of interest with a solution containing one or more alkylamine compounds. In a particular embodiment, the active ingredients are amphipathic, charged amines or amine oxides coupled to saturated hydrocarbon chains of varying lengths. In a preferred embodiment, the one or more active ingredients used are selected from the group consisting of dimethyldecylamine, dimethyltridecylamine, dimethylundecylamine, dimethyldidecylamine, dimethyltetradecylamine, dimethylhexadecylamine, dimethyldecylamineoxide, dimethylundecylamineoxide, dimethyldidecylamineoxide, dimethytetradecylamineoxide and dimethyltridecylamineoxide. These compounds may be used at concentrations ranging from 0.001% up to their solubility limit in the given solution. Preferably, the concentration of the active ingredients ranges from 0.005% to 5%, 0.1% to 2%, or is approximately 0.5% of the total solution (weight basis).
The pH of the solution can range from pH 2 to pH 12. Preferably, the solution is at a pH ranging from pH 5.0 up to pH 8.0. More preferably, the pH ranges from pH 5.5 to 7.4, from pH 6 to 7.4, from pH 7.0 to 7.4, or is approximately pH 7.2.
The inactivation of the viral contaminants with the active ingredient of the invention can be carried out at a temperature of from about 2xc2x0 C. to about 50xc2x0 C. Preferably, the temperature is from about 2xc2x0 C. to about 30xc2x0 C., 2xc2x0 C. to about 20xc2x0 C., 2xc2x0 C. to about 10xc2x0 C., about 4xc2x0 C., about 25xc2x0 C., or at room temperature.
The biological source material may be blood plasma, other biological tissues or a recombinant source material such as transformed or transfected xe2x80x9chost cellsxe2x80x9d. xe2x80x9cHost cellsxe2x80x9d are cells containing a biomolecule of interest. A xe2x80x9cbiomolecule of interestxe2x80x9d is any biomolecule present in the biological source material or the host cells that one desires to isolate, purify, or formulate for subsequent processing or application. The biological source material may be blood, blood plasma, animal tissues, plant tissues or recombinant host cells or host cell extracts. The host cells may be of any type, preferably mammalian, bacterial, yeast, fungal, plant, avian, insect, or reptilian.